Changes in rat spermatozoa function after cooling, cryopreservation and centrifugation processes
Changes in rat spermatozoa function after cooling, cryopreservation and centrifugation processes
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DOI：
10.1016/j.cryobiol.2012.06.006
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发表时间：
2012-12-01
期刊：
影响因子：
2.7
通讯作者：
Agca, Yuksel
中科院分区：
文献类型：
--
作者：
Kim, Suhee;Agca, Cansu;Agca, Yuksel
大鼠精子低温保存是为生物医学研究保存有价值菌种的有效方法，大鼠精子的处理是成功低温保存的关键。本研究的目的是评估大鼠精子在冷冻保存和离心过程中功能的变化。对附睾大鼠精子进行冷却和冻融处理，比较最小离心力（200 xg）前后的运动性、质膜完整性（PMI）、线粒体膜电位（MMP）和活性氧（ROS）。低温保存降低了精子活力。PMI和MMP （P < 0.05）。与新鲜精子相比，冷冻活精子的基础（不含ROS诱导剂，叔丁基过氧化氢[TBHP]处理）和刺激ROS（有TBHP处理）均增加（P < 0.01），新鲜、冷却和冻融精子对TBHP的敏感性相同。离心降低了冷冻解冻精子的活力和PMI （P < 0.05）。离心法降低了所有精子的基础ROS (P < 0.01)，而离心法提高了低温活精子对thbhp的敏感性（P < 0.05），表明低温活精子的ROS增加倍数更高，thbhp降低了活精子的存活率（P < 0.05）。冷却过程是ROS生成的主要步骤，伴随着精子活力、PMI和MMP的丧失。离心法影响冷冻精子的功能。这些数据表明，离心使大鼠精子容易受到外部活性氧源的影响，特别是在冷却过程中。因此，在冷冻保存和解冻后使用大鼠附睾精子时，应考虑防止ROS损伤和尽量减少离心。Elsevier Inc.出版。
Rat sperm cryopreservation is an effective method of archiving valuable strains for biomedical research and handling of rat spermatozoa is very important for successful cryopreservation. The aim of this study was to evaluate changes in rat sperm function during cryopreservation and centrifugation. Epididymal rat spermatozoa were subjected to cooling and freezing-thawing processes and then motility, plasma membrane integrity (PMI), mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) were compared before and after minimum centrifugation force (200 xg). Cryopreservation decreased sperm motility. PMI, and MMP (P < 0.05). Basal (without ROS inducer, tert-butyl hydroperoxide [TBHP] treatment) and stimulated ROS (with TBHP treatment) were increased in viable cooled spermatozoa compared to viable fresh spermatozoa (P < 0.01), with equal susceptibility to TBHP among fresh, cooled, and frozen-thawed spermatozoa. Centrifugation decreased motility and PMI of frozen-thawed spermatozoa (P < 0.05). Centrifugation decreased basal ROS of all spermatozoa (P < 0.01), while it led to higher susceptibility to TBHP in viable cooled spermatozoa, showing higher increased fold in ROS and decreased rate in viability by TBHP in viable cooled spermatozoa (P < 0.05). Cooling process was the major step of ROS generation, with loss in sperm motility, PMI, and MMP. Centrifugation affected function of cryopreserved spermatozoa. These data suggest that centrifugation makes rat spermatozoa susceptible to external ROS source, in particular during cooling process. Thus, protection from ROS damage and minimizing centrifugation should be considered during cryopreservation and post-thaw use of cryopreserved epididymal rat spermatozoa. Published by Elsevier Inc.